Physical Non-Viral Gene Delivery Methods for Tissue Engineering

Abstract

The integration of gene therapy into tissue engineering to control differentiation and direct tissue formation is not a new concept; however, successful delivery of nucleic acids into primary cells, progenitor cells, and stem cells has proven exceptionally challenging. Viral vectors are generally highly effective at delivering nucleic acids to a variety of cell populations, both dividing and non-dividing, yet these viral vectors are marred by significant safety concerns. Non-viral vectors are preferred for gene therapy, despite lower transfection efficiencies, and possess many customizable attributes that are desirable for tissue engineering applications. However, there is no single non-viral gene delivery strategy that “fits-all” cell types and tissues. Thus, there is a compelling opportunity to examine different non-viral vectors, especially physical vectors, and compare their relative degrees of success. This review examines the advantages and disadvantages of physical non-viral methods (i.e., microinjection, ballistic gene delivery, electroporation, sonoporation, laser irradiation, magnetofection, and electric field-induced molecular vibration), with particular attention given to electroporation because of its versatility, with further special emphasis on Nucleofection™. In addition, attributes of cellular character that can be used to improve differentiation strategies are examined for tissue engineering applications. Ultimately, electroporation exhibits a high transfection efficiency in many cell types, which is highly desirable for tissue engineering applications, but electroporation and other physical non-viral gene delivery methods are still limited by poor cell viability. Overcoming the challenge of poor cell viability in highly efficient physical non-viral techniques is the key to using gene delivery to enhance tissue engineering applications.

Experimental approaches to vascularisation within tissue engineering constructs

“…[261] Nonetheless, physical delivery methods are not free from drawbacks, for example, they may trigger apoptosis, reduce cell viability and can negatively affect cell phenotype. [262] Alternatively, in chemical methods, cationic lipids, cationic polymers, cell-penetrating peptides, and inorganic nanoparticles can be used to enhance DNA delivery efficiency to target cells, [263] although large doses of chemical vector to achieve high DNA transfection efficiency can cause cytotoxicity. [264] A number of studies have used viral and nonviral vectors to evaluate the effect of gene therapy in vascular tissue engineering.…”

MagRET Nanoparticles: An Iron Oxide Nanocomposite Platform for Gene Silencing from MicroRNAs to Long Noncoding RNAs

“…32 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 BxPC-3 cells with MagRET NPs complexed with a mir-21 inhibitor led to a significant downregulation of 89±5% at a concentration of 100 nM ( Figure 6A). …”

“…So far, siRNA delivery to cells in suspension can be achieved almost exclusively by electroporation which is expensive, cytotoxic, and cannot be considered for in vivo applications 32. CMK cells were established from a Down syndrome patient with acute megakaryoblastic leukemia 33.…”

Copolymer micelles function as pH-responsive nanocarriers to enhance the cytotoxicity of a HER2 aptamer in HER2-positive breast cancer cells

“…[16][17][18][19] Viral vectors, such as retrovirus or adenovirus-based systems, have high transduction efficiencies. However, their application is limited by high production costs and biosafety and immunogenicity concerns.…”